RU2060010C1 - Fish food - Google Patents

Abstract

FIELD: fish breeding, veterinary. SUBSTANCE: fish food involves the milled food components and the medicinal agent - a compound of the general formula

where R - alkanoyl-group, benzoyl-group with one or more substituents taken from the group consisting of the lower alkyls and halogen atoms, phenylacetyl-group or hexane carbonyl-group. Food containing the compound of the general formula

Description

O где R алконоильная группа; бензоильная группа, возможно имеющая одну или более групп-заместителей, выбранных из группы, состоящей из низших алкилов и галоген-атомов; фенилацетильная группа; или циклогексан-карбонильная группа.The invention relates to fish feed containing ground food components and a drug [1]
The food according to the invention as a medicine contains a compound of the following General formula (I):

O where R is an alkonoyl group; a benzoyl group, possibly having one or more substituent groups selected from the group consisting of lower alkyls and halogen atoms; phenylacetyl group; or cyclohexane carbonyl group.

 In recent years, significant successes have been achieved in the fight (when breeding freshwater and marine fish) with concomitant outbreaks of various fish diseases. To prevent and treat certain fish diseases, for example pseudotuberculosis in the yellowtail, antibacterial drugs such as ampicillin, oxolinic acid and the like are often used. However, the usual use of these antibacterial drugs presents a problem in that pathogenic bacteria resistant to these antibacterial drugs appear with an increase in the frequency of use.

 Therefore, there is an urgent need for the creation of new antibacterial drugs for the prevention and treatment of diseases in fish that would not have the unfavorable immunity of traditional drugs and would be active against such refractory microorganisms.

 To solve this problem, the applicants intended to use bicosamycin (the same substance as the substance WS-4545, which is produced by some microorganisms of the genus streptomyces, Japanese Patent Application Publication No. 29158, 1973), but found that this substance did not possess satisfactory digestibility from the intestinal tract.

 Meanwhile, some esters synthesized to improve the absorption of bicosamycin after oral administration (the same substances as the acylated substance WS-4545, Japanese Patent Application Laid-Open No. 39497, 1973) met this requirement (Fhe Iournal of Antibiotics, t. XXY, N 10, pp. 576-581), however, refrained from using such bicosamycin esters for fish for the following reason. Thus, medications for fish are usually administered in the form of raw minced fish, but it is usually believed that if bicosamycin is introduced to fish in this way, especially yellowtail, this ether will be hydrolyzed back to bicosamycin under the influence of esterase found in minced fish so that the goal of improving the oral absorption of the drug will not be achieved. (Indeed, when a well-known ampicillin ester, namely bicampicillin, is placed in minced fish before use, it decomposes in minced meat).

 Under these conditions, they tried to experimentally inject fish with bicosamycin ester, compound (I) in the aforementioned manner, and unexpectedly found that this ester practically did not decompose in minced fish, but it was well absorbed by the blood and remained there with a high concentration for a long time. This discovery was followed by research, the results of which are set forth in this invention.

 The aim of the invention is to overcome the above problems, and it is directed to fish food intended for the prevention and treatment of diseases in fish and containing compound (I) as an active ingredient.

 In accordance with this invention, the compound (I) used, as described above, is a substantially known compound. In addition, bicosamycin, the starting material for this compound (I), is known as an antibiotic produced by Streptomyces sapporonensis ATCC 21532, as described in Japanese Patent Application Publication No. 29158, 1973, referred to above.

 The producer strain mentioned above.

 Depository: American Type Culture Collection.

 Address: Parklawn Drive 12301, Rockville, ellaryland, USA.

 Date of deposit: April 21, 1970.

 Depository N: ATCC 21532.

 Speaking of compound (I), preferred examples of said alkanoyl group include acetyl, propionyl, butyryl, isobutyryl, valeryl, isovaleryl, octanoyl, pilmitoyl and so on; preferred alkyls of said lower alkyl group are methyl, ethyl, propyl and so on; and preferred halogens are chlorine, bromine and iodine.

 The following is a partial list of representative representatives of compound (I) that may be used in this invention.

(I)

1 -CO

2 -CO

3 -CO

CH₃ 4 -CO

CH₃ 5 -COCH

6 -COCH

7 -COCH₂CH

8 -CO

C₁ 9 CO (CH₂)₁₄ CH₃ 10 COCH₂CH₂CH₃
Рыбы и их заболевания, для которых применим корм по изобретению, включают среди прочих псевдотуберкулез (Pasteurella piscicida) желтохвоста и каранга, вибриоз (Vibrio anguillarum) желтохвоста, айю и морского карася, эдвардсиллез (Edwardsiella tarda) японского угря, плоской рыбы и морского карася и фурункулез (Aeromonas salmonicida) радужной форели.

(I)

1 -CO

2 -CO

3 -CO

CH ₃ 4 -CO

CH ₃ 5 -COCH

6 -COCH

7 -COCH ₂ CH

8 -CO

C ₁ 9 CO (CH ₂ ) ₁₄ CH ₃ 10 COCH ₂ CH ₂ CH ₃
Fish and their diseases for which the food according to the invention is applicable include, among others, pseudotuberculosis (Pasteurella piscicida) of yellowtail and Karanha, vibriosis (Vibrio anguillarum) of yellowtail, ayu and crucian carp, edwardsillosis (Edwardsiella tarda) of Japanese eel, flat fish and sea fish furunculosis (Aeromonas salmonicida) rainbow trout.

 The food according to the invention can be prepared by processing compound (I) into a dosage form such as powder, dust, micronized granules, granules, micronized granules, tablets, liquids, pills or syrups with or without a solid, semi-solid or liquid carrier, and food components to formulate a feed containing compound (I) or the specified dosage form.

 The medium includes, among others, raw minced fish (e.g. finely chopped mackerel, sardines, kremerievye, mackerel shrimp, pollock, squid and so on), prescription food based on fishmeal, soybean meal, yeast, wheat flour, vitamins and so on, and also conventional carriers such as lactose, sucrose, glucose, starch, talc, acid clay and so on.

 In addition, emulsifiers, dispersants, gelling agents, binders and so on can be added in appropriate amounts.

 Foods containing compound (I) can be used to prevent diseases and treat diseases in yellowtail, karanga, ayu, carp, crucian carp, Japanese eel, flat fish, rainbow trout and so on. For example, for the prevention and treatment of pseudotuberculosis in yellowtail, the most preferred therapeutic effect, given the advantage in the stability of compound (I) in minced fish, is achieved by adding compound (I) in the form of a powder or fine granules pre-mixed with the specified medium to raw minced fish or to a mixture of such raw minced fish and prescription food, and the introduction of the whole mixture either in the form as it is, or pre-compressed in the form of granules or wet granules.

 The dosage and duration of feeding for the treatment of diseases in fish depends on the type and age of the fish, water temperature, severity of the disease, and so on. For example, for the prevention and treatment of pseudotuberculosis in the yellowtail, typically 1 to 50 mg orally can be administered per compound (I) per day per 1 kg of fish weight for 3-10 days.

 Food containing compound (I) does not exhibit adverse immunity of traditional drugs, is active against immune strains of microorganisms, and is stable in the form of raw minced fish. Therefore, when a feed in the form of a mixture with raw minced fish is introduced to the fish, this feed guarantees with sufficient reliability in the blood of the fish a high concentration of bicosamycin for a long period of time. The following test examples are intended to illustrate the effectiveness of this invention.

 Test 1 (temporary dependence of the concentration of bicosamycin in the blood of the yellowtail).

 The active compound was mixed with crushed karang and administered to the yellowtail freely, in an affordable manner, as a single dose of 50 mg / kg. 6, 9, 12, 24, 36, 48, and 72 hours after administration, blood was taken from the heart.

 Then, the concentration of bicosamycin in the blood was determined by means of a biological test using E. coli BS-10 as a test microorganism.

 The results are shown in table 1.

 Test 2 (stability in enzyme solutions 1).

 Each test drug was dissolved in water to a final concentration of 2000 μg / ml.

 Then, solutions of each drug (0.1 ml portions) were added to various enzyme solutions (0.9 ml portions).

 Enzyme solutions were prepared as follows.

 Deionized water: control for lack of enzyme.

 Yellowtail plasma: blood was taken from the yellowtail heart into a heparinized syringe, centrifuged for 15 minutes at 3000 rpm and plasma was obtained.

 Yellowtail liver homogenate: a liver was taken from a lively grown yellowtail, crushed and homogenized.

 Yellowtail gut homogenate: a live gut yellowtail was taken from the gut, crushed and homogenized.

 Pork esterase: Commercially available pork esterase (Sigma) was diluted with a 50-fold volume of water.

 Chopped mackerel: they bought fresh mackerel, chopped all the fish and passed through a meat grinder. Minced fish was homogenized with 2 volumes of water, centrifuged for 15 min at 3000 rpm and the supernatant was used as an enzyme solution.

 Chopped sardine: obtained in the same way as chopped mackerel.

 Chopped mackerel: obtained in the same way as chopped mackerel.

Drug solution was mixed with enzyme solution and reacted at ²⁵ ° C for 2 hours.

 In order to quench the enzymatic reaction, 1.0 ml of methanol was added.

 The reaction mixture was carefully mixed and centrifuged for 5 min at 6400 rpm. A paper disk was immersed in the supernatant and the concentration of ampicillin, cefteram (a related compound to pivoxil ceftheramine) or bicosamycin was determined by means of a bioassay using E. coli BS-10 as the test strain.

 The results are shown in table.2. The numerical data presented are the rates of conversion of bacampicillin to ampicillin, pivoxil ceftheram to its related compound (Journal of. Japan Society of chemotherapy, v. 34, S-2, pp. 44-60, 1986) and the compounds of this invention to bicosamycin.

 Test 3 (stability in enzyme solutions 2).

 Each test drug was dissolved in water to a final concentration of 1000 μg / ml.

 Then, 0.04 ml of each drug solution was added to 0.36 ml portions of various enzyme solutions.

 Enzyme solutions were prepared as follows.

 Deionized Water: Enzyme Control

 Pork esterase: Commercially available pork esterase (Sigma) was diluted with a 50-fold volume of water.

 Raw minced fish: fresh mackerel and sardine were bought and equal proportions of the corresponding whole fish were crushed and mixed. This mixture was passed through a meat grinder, homogenized with 2 volumes of water and centrifuged for 15 min at 3000 rpm. The supernatant was used as the enzyme solution.

 Yellowtail serum: blood was taken from the yellowtail's heart into a syringe, kept at room temperature for 1 h, and then centrifuged for 15 min at 3000 rpm to obtain serum.

 Rainbow trout plasma: blood was taken from the heart of the rainbow trout into a heparinized syringe, centrifuged for 15 min at 3000 rpm and plasma was obtained.

Drug solution was mixed with enzyme solution and reacted at 30 ^{° C} for 3 hours.

 In order to quench the enzymatic reaction, 0.4 ml of methanol was added.

 After stirring carefully, the reaction mixture was centrifuged at 64,000 rpm for 5 minutes. A paper disk was immersed in the supernatant and bicosamycin concentration was determined by bioassay using E. coli BS-10 as the test strain.

 The results are shown in table.3. The figures given are the conversion rates of the compounds of this invention to bicosamycin.

 Test 4 (comparison with traditional drugs).

 Yellow tails weighing about 200 g each, which had developed pseudotuberculosis in a large space, were divided into 7 groups of 200 fish and were treated according to the following regimen for the use of the drug. Group I: a control group that was not given drugs; group II: ampicillin 20 mg / kg; group III: oxolinic acid 30 mg / kg; group IV: compound I 20 mg / kg; group V: compound I 40 mg / kg; group VI: compound 2 20 mg / kg; Group VII: Compound 2 40 mg / kg. The total weight of the fish in each group was determined. In each group of fish, they were not fed for one day, and then they were given once a day for 5 consecutive days a mixture of the yellow tail and prescription food mixture (10% in lactose) as an additional mixture to the crushed cremerias. Every day, in the morning hours, dead fish were pulled out and the number of dead fish was recorded. In addition, to confirm that death was due to pseudotuberculosis, all dead fish were autopsied.

 Daily, in the morning hours, the temperature of the water was measured and recorded.

 The results are shown in table 4.

 Test 5 (comparison with bicosamycin).

 The yellow tails weighing about 200 g each, which had developed pseudotuberculosis in a large space, were divided into 4 groups of 300-320 fish and treated according to the following regimen of drug use. Group I: a control group that was not given drugs; group II: bicosamycin 20 mg / kg; group III: compound I 20 mg / kg; Group IV: Compound 2 20 mg / kg. The total weight of the fish in each group was determined. The fish in each group were not fed for one day, and then they were given once a day for 5 consecutive days a mixture of prescription food for yellowtail and the corresponding drug (10% in lactose) as an additional mixture to the crushed cremerii. Every day, in the morning hours, dead fish were pulled out and the number of dead fish was recorded. In addition, to confirm that death was due to pseudotuberculosis, all dead fish were autopsied.

 Daily in the morning hours, the temperature of the water was measured and recorded.

 The results are shown in table 5.

 PRI me R 1. The powder is obtained by mixing 6.7 wt.h. compounds I (according to the effectiveness corresponds to 5 parts by weight of bicosamycin) with 93.3 parts by weight lactose free.

 Using 100 g of this powder as a unit of daily dose, about 20,000 yellow tails are fed (the weight of one yellow tail was about 50 g, and the total weight of all fish is 1 ton) with a mixture containing this daily dose and 200 kg of cremeria mince for 5 consecutive days . The above nutritional regimen achieves the goal of preventing and treating yellowtail pseudotuberculosis.

 PRI me R 2. The powder is obtained by mixing 6.7 wt.h. compounds I (according to the effectiveness corresponds to 5 parts by weight of bicosamycin) with 93.3 parts by weight lactose free.

 About 15,000 yellow tails (the weight of one yellow tail is about 200 g, and the total weight of all fish is 3 tons) produce wet granules (cylindrical granules obtained by mixing minced sardines with powdered fish meal formulated feed in a ratio of 6: 4 s for 5 consecutive days) subsequent mechanical granulation of this mixture), to which 300 g of the above powder were added. This dietary regimen achieves the goal of preventing and treating pseudotuberculosis in yellow tails.

 Based on the foregoing, the feed according to the invention can be used as a prophylactic / therapeutic agent, in particular a therapeutic agent for the treatment of fish diseases and, thus, to increase fish production.

Claims (1)

Fish food containing crushed food components and a medicinal product, characterized in that as a medicine it contains a compound of the general formula

R is an alkonoyl group, a benzoyl group optionally having one or more substituent groups selected from the group consisting of lower alkyls and halogen atoms, a phenylacetyl group or a cyclohexanecarbonyl group.

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